Electrochemical detection of PCR amplicons of Escherichia coli genome based on DNA nanostructural probes and polyHRP enzyme
Wen, YL; Wang, LL; Xu, L; Li, LY; Ren, SZ; Cao, CM; Jia, NQ; Aldalbahi, A; Song, SP; Shi, JY; Xia, JY; Liu, G; Zuo, XL; Liu, G (reprint author), Shanghai Inst Measurement & Testing Technol, Div Chem & Ionizing Radiat Measurement Technol, Shanghai 201203, Peoples R China.; Zuo, XL (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Div Phys Biol, Shanghai 201800, Peoples R China.; Aldalbahi, A (reprint author), King Saud Univ, Dept Chem, Riyadh 11451, Saudi Arabia.
2016
发表期刊ANALYST
ISSN0003-2654
卷号141期号:18页码:5304-5310
文章类型期刊论文
摘要Fast, portable and sensitive analysis of E. coli is becoming an important challenge in many critical fields (e.g., food safety, environmental monitoring and clinical diagnosis). Thus, electrochemical biosensing of PCR amplicons from the bacterial genome has attracted reasonable research attention. In this work, we utilized a 3D DNA tetrahedral probe to establish a "sandwich-type" electrochemical DNA biosensor for sensitive and specific analysis of a 250 bp unpurified PCR amplicon from the uidA gene of the E. coli genome. Asymmetric PCR was used to produce single-stranded PCR products. Streptavidin-polyHRP80 was employed to improve the signal gain during electrochemical detection. We optimized important experimental conditions for DNA sensing, including the streptavidin-polyHRP, the signal probe and the ion strength. Finally, we achieved a remarkable sensitivity of 10 fM synthetic DNA target, and successfully performed the analysis of PCR amplicons from as low as 0.2 pg mu L-1 of E. coli genome. Compared with traditional single stranded DNA (ssDNA) probe based detection, our present work demonstrated 3 orders of magnitude improvement in sensitivity. In addition, our electrochemical DNA biosensor was 4 orders of magnitude more sensitive than normal electrophoretic analysis of PCR products. Our work made important progress in DNA nanostructured probe-based biosensors toward application in real applications.
DOI10.1039/c6an01435f
收录类别SCI
语种英语
WOS记录号WOS:000382698300012
引用统计
被引频次:6[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.sinap.ac.cn/handle/331007/26493
专题中科院上海应用物理研究所2011-2018年
通讯作者Liu, G (reprint author), Shanghai Inst Measurement & Testing Technol, Div Chem & Ionizing Radiat Measurement Technol, Shanghai 201203, Peoples R China.; Zuo, XL (reprint author), Chinese Acad Sci, Shanghai Inst Appl Phys, Div Phys Biol, Shanghai 201800, Peoples R China.; Aldalbahi, A (reprint author), King Saud Univ, Dept Chem, Riyadh 11451, Saudi Arabia.
推荐引用方式
GB/T 7714
Wen, YL,Wang, LL,Xu, L,et al. Electrochemical detection of PCR amplicons of Escherichia coli genome based on DNA nanostructural probes and polyHRP enzyme[J]. ANALYST,2016,141(18):5304-5310.
APA Wen, YL.,Wang, LL.,Xu, L.,Li, LY.,Ren, SZ.,...&Aldalbahi, A .(2016).Electrochemical detection of PCR amplicons of Escherichia coli genome based on DNA nanostructural probes and polyHRP enzyme.ANALYST,141(18),5304-5310.
MLA Wen, YL,et al."Electrochemical detection of PCR amplicons of Escherichia coli genome based on DNA nanostructural probes and polyHRP enzyme".ANALYST 141.18(2016):5304-5310.
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